Hatchery-reared salmonid fish routinely encounter stress due to handling, barging, tagging, and overcrowding. It has been demonstrated that there exists a direct correlation between stress and transient immune suppression which can last for many days in fish. Epizootic viral infections routinely appear in hatcheries and can have a devastating effect on the fish population. The major viral pathogens in salmon and trout are the fish rhabdovirus, infectious hematopoietic necrosis virus (IHNV), and the fish birnavirus, infectious pancreatic necrosis virus (IPNV). Vaccines for these viral pathogens are under investigation; however, the fish immune system becomes virtually nonresponsive during episodes of immune suppression. It was necessary to develop a nonantibody mediated, nonimmune method for preventing viral infections. An interferon-like substance has been described for fish which possesses antiviral activity against both IHNV and IPNV. Since interferon administered to cattle has been very effective against vesicular stomatitis virus, a cattle rhabdovirus, an examination of interferon-like activity in fish was initiated. We report here the establishment of in vitro interferon assays. In addition, the salmonid genome contains a multigene family of p-interferon-like genes, much like those in the bovine, equine and porcine genome. The rainbow trout interferon-like genes were found to be inducible in a manner which parallels those seen with bovine and human interferons. In addition to the multigene interferon-like family, it was found that rainbow trout also contain a retroposon multigene family. Retroposons are repetitive elements which appear to have arisen by a reverse transcription event. Two Ll like repetitive elements have been cloned, one of which contains a Drosophila retroposon polymerase sequences never before described for salmonid fish. A number of retroviruses have been described in fish including the walleye dermal sarcoma virus and the Atlantic salmon swimbladder sarcoma virus. Interferon shows prophylactic promise both in vivo and in vitro, against the human retrovirus, HIV. Therefore, research into fish interferon may be even more important if it demonstrates not only anti- IHNV and anti-IPNV, but also anti-fish retrovirus properties. / Graduation date: 1993

Primary Epstein-Barr virus (EBV) infection in childhood is typically asymptomatic, but infection later in life results in a mononucleosis with the illness severity ranging from asymptomatic to requiring hospitalisation. The illness is generally short-lived (four to six weeks following onset of symptoms), but persistent disabling symptoms (lasting up to 6 months or longer) are well described in around ten percent of individuals. The aim of this work was to characterise immunological and virological parameters in the peripheral blood, which correlate with persistence of symptoms following EBV-induced mononucleosis. Subjects were recruited prospectively following confirmed primary EBV infection, to allow blood samples and clinical data to be collected at multiple timepoints (baseline, 2 weeks, 4 weeks, 3 months and at least 12 months later). Subjects with 6 months or more of disabling symptoms were defined as ???cases??? with control subjects being those whose illness resolved within 6 weeks of enrolment. Cases were compared with control subjects in terms of: cellular EBV viral load in the peripheral blood by PCR; development of antibodies against EBV VCA (IgG and IgM) and EBNA-1 (IgG) by ELISA; proportions of peripheral blood leucocyte subsets and their activation status by flow cytometry; the magnitude, kinetics of development, and breadth of the CD8+ cytotoxic cell response by interferon-?? Elispot; cytokine levels in serum, and production by peripheral blood mononuclear cells ex vivo; and gene expression patterns in peripheral blood mononuclear cells by microarray. With the exception of gene expression, none of these parameters correlated with early resolution of symptoms or predicted clinical outcome following primary EBV infection. Antibody patterns suggest a tendency to Th2 type immune response may be associated with persistent illness. Preliminary analysis of the gene expression studies indicates that there are many genes involved in this complex disease requiring further investigation. Persistent illness following EBV infection is not associated with uncontrolled viral replication, or chronic immune activation due to an aberrant primary immune response.

The circovirus Beak and feather disease virus (BFDV) causes psittacine beak and feather disease (PBFD) that is characterised by a chronic disease process associated with feather abnormalities, beak deformities and eventual death in various species of birds in the order Psittaciformes. This disease is seen in captive and wild psittacine species in Australia and several other countries and is a significant threat to the survival of some endangered psittacine species. This thesis reports on genetic studies that have furthered the understanding of the diversity of BFDV present within Australia. These studies have optimised methods of detecting BFDV. They have also resulted in the production of an immunogenic and antigenic recombinant BFDV Capsid protein that could lead to alternate methods of producing viral antigen for serological tests and the development of a BFDV vaccine. To assess the optimal method of the detection of BFDV infection, feather and blood samples were submitted by referring veterinarians throughout Australia from psittacine birds tentatively diagnosed with PBFD or with a history of being in contact with PBFD-affected birds. These samples were examined by 3 procedures commonly used to detect BFDV infection: a polymerase chain reaction (PCR) assay and haemagglutination (HA) for the detection of virus, and haemagglutination inhibition (HI) tests for the detection of virus antibody in response to infection. Of the samples examined from 623 psittacine birds, the prevalence of BFDV DNA in feather samples detected by PCR was 18.85%. There was a strong correlation between PCR and HA testing of feather samples, although possible false-positive and false-negative PCR and HA results were obtained in some samples. Of the 143 birds that were PCR feather-positive only 2 had detectable HI antibody and these birds were also HA feather-negative, which suggests that they were developing immunity to recent infection. All birds with HI antibody were feather HA negative. Despite the rare occurrence of PBFD in cockatiels (Nymphicus hollandicus), 2 of the 13 samples collected from this species were PCR and HA positive indicating that this species can be infected with BFDV. Three studies were undertaken to further our understanding of the genetics of BFDV in Australian avifauna: sequence analysis of the BFDV detected in a grey cockatiel (Nymphicus hollandicus), a species normally considered resistant to infection with BFDV; analysis of the genome of BFDV present in lorikeets (Trichoglossus sp.) in Australia; and analysis of the genome of BFDV detected in endangered swift parrots (Lathamus discolor). Sequence analysis of the entire genome of the cockatiel BFDV isolate revealed that it clustered phylogenetically with 2 other viruses, one from a sulphur crested cockatoo (SCC1-AUS) and one from a Major Mitchell cockatoo (MMC-AUS), which suggests that this isolate from the grey cockatiel was not a cockatiel-specific biotype. Phylogenetic analysis of the ORF V1 of BFDV detected in 7 lorikeets demonstrated these 7 isolates clustered phylogenetically with other BFDV isolates obtained from Loriidae species elsewhere in the world and confirmed the presence of a loriid-specific genotype. Phylogenetic analysis of the sequence data generated from ORF V1 of virus detected in 2 endangered swift parrots provided evidence they were also infected with BFDV genotypes derived from other species of birds, one isolate clustering with viruses from a Loriidae genotype and the other with isolates derived from species of Cacatuidae and Psittacidae. As part of this research, a baculovirus expression system was successfully developed for the production of recombinant BFDV Capsid protein. Inoculation of this protein into chickens resulted in the development of HI antibody, which demonstrated its immunogenicity. When used as an antigen in HI tests it detected antibody in virus-infected birds, which demonstrated its antigenicity. This protein offers potential application as an antigen for the development of serological tests and as an immunogen for incorporation into vaccines for control of PBFD.

Thesis (Ph. D.)--University of Adelaide, Dept. of Plant Pathology, 1985. / Includes bibliographical references (leaves 119-127).

Thesis (Ph.D.) -- McMaster University, 1998. / Includes bibliographical references (leaves 333-365). Also available via World Wide Web.

Thesis (Ph. D.)--University of Hong Kong, 2008. / Also available in print.

Diss. (sammanfattning) Uppsala : Uppsala universitet, 2005. / Härtill 5 uppsatser.

Thesis (Ph. D.)--University of Adelaide, 1993. / Includes bibliographical references (leaves 102-125).

Thesis (Ph. D.)--University of Adelaide, Dept. of Crop Protection, 1997. / Includes bibliographical references (leaves 133-152).

Thesis (M. Phil.)--University of Hong Kong, 2008. / Includes bibliographical references (leaf 102-115) Also available in print.